BIOLOGI MOLEKULER

ONKOLOGI
dr. Henny E. S. Ompusunggu,M.Biomed
Dept. Biologi Sel dan Molekuler, Fakultas
Kedokteran Universitas HKBP Nommensen,
Medan

BIOLOGI MOLEKULER ONKOLOGI

Onkos (Yunani ) = tumor (massa)

Kanker merupakan kelainan genetik

kompleks yang melibatkan kelainan
struktural dan kelainan ekspresi gen

Penyebab: perubahan protein yang

mengkode protoonkogen dan tumor
supresor

Multiple genetic changes underlie the

development of cancer

Chromosomes
1
mutation

Normal
cell

4
2
3
mutations mutations mutations

Malignant
cell

Four or more somatic mutations are usually

required to produce a cancer cell

Klasifikasi tumor menurut sel embrionik

asalnya :
1. Karsinoma
>80% tumor
Berasal dari jaringan endodermal/ektodermal
kulit, lapisan epitel organ internal, dan
kelenjar.

2. Kanker sel darah

Hematopoietic cells Leukemia dan
Limfoma

3. Sarkoma
Jaringan ikat jaringan mesodermal tulang,
lemak dan tulang rawan

SIKLUS SEL
Siklus sel merupakan fungsi sel
yang paling mendasar berupa
duplikasi akurat sejumlah besar DNA
di dalam kromosom, dan kemudian
memisahkan hasil duplikasi tersebut
hingga terjadi dua sel baru yang
identik.
Siklus sel yang berlangsung kontinu
dan berulang (siklik), disebut

FASE G (Gap)
Fase sintesis zat yang diperlukan pada fase
berikutnya
G1: Check point, Reparasi DNA yang
bermutasi. Bila tidak dapat direparasi maka
akan dilakukan Apoptosis (oleh enz.
Kaspase)
Interval Fase G1 sangat bervariasi antara 6
jam hingga beberapa hari.
G2: persiapan memasuki fase pembelahan
sel
Interval fase G2 sekitar 2 jam

Fase S (Sintesis)
Tahap terjadinya replikasi (sintesis
DNA baru)
Membutuhkan waktu sekitar 8 jam
Hasil replikasi kromosom yang telah
utuh akan memasuki fase M.

DNA Replication
A
T
DNA
unzips

A
G

New strands
formed

C
T

A
G

G
A
T
C

semi-conservative
2 daughter cells

Replikasi DNA berlangsung pada kedua

helix dengan arah yang berlawanan

DNA Replication
Replication fork : leading strand and lagging
strand
DNA synthesized in the 5 3
The 5-3 synthesis of the leading strand is
continuous.
The lagging strand is also synthesized in the 53 direction but in small segments
This segments referred to as Okazaki fragments
Okazaki fragments has 100 200 nucleotides
DNA ligase joined the Okazaki fragments.

Fase M (Mitosis/Meiosis)
Merupakan tahap pembelahan sel.
Interval waktu fase M kurang lebih 1
jam.
Pada mitosis, sel membelah dirinya
membentuk dua sel yang terpisah,
sementara pada meiosis sel
membelah menjadi 4 sel.
Interfase : merupakan sebuah jedah
panjang antara satu mitosis dengan
yang lain (Jedah tersebut termasuk

Fase M
(Mitosis/Meiosis
)

BIOLOGI MOLEKULER ONKOLOGI

PROTOONKOGEN :
Gen yang normal memicu pembelahan sel yang
normal (proliferasi & diferensiasi), cth: ras, erb
B, src, bcl, myc, HER2.
ONKOGEN :
Protoonkogen yang bermutasi memicu
pembelahan sel yang tidak terkontrol (menjadi
tumor)
TUMORSUPPRESSOR GEN :
Gen yang normal menghambat pembelahan sel.
Bila bermutasi akan menyebabkan pembelahan
sel menjadi tidak terkontrol, cth: Rb, p53, PTEN,
BRCA.

Produk Protoonkogen
SIS

ABL

SRC
RAS

FMS

Inti Sel
FOS
MYC
JUN

O
rg
an
el
la
MOS

ERB-B1

FMS

ONCOGENE
Proto-oncogene Mutation Oncogene
Oncogenes :

promote cell proliferation

dominant & highly conserved

types:

viral oncogenes [v-oncs]

cellular oncogenes [c-oncs]

ONCOGENE FAMILY
Classification of Oncogenes
A. Secreted Growth Factors
c-sis, hst

B. Cell Surface Receptors

erb B, fms, ret, trk, fes, fms

C. Intracellular Transducers
c-src, c-abl, mst, ras

D. DNA-binding Nuclear Proteins

myc, jun, fos

E. Regulators of the Cell Cycle

bcl, bax, bad

Components of
signal transduction
pathways

Molecular Biologi of Cancer. Burgers Medicinal Chemistry and Drug Discovery Sixth
Edition, Volume 5: Chemotherapeutic Agents. Edited by Donald J. Abraham. ISBN 0-47137031-2 2003 John Wiley&Sons, Inc.

SIGNAL
TRANSDUCTION

Cell Signaling

TUMOR SUPPRESSOR GENE

TS Genes

inhibit growth and multiplication of mutated cells

prevent neoplastic transformation

recessive & highly conserved

Classification of TS genes
A. Cell Adhesion Molecules
APC, DCC

B. Regulators of the Cell Cycle

RB1, Tp53

APOPTOSIS
APOPTOSIS programmed cell
death

Balance between proliferation and

apoptosis is critical in determining
growth or regression

Molecular Biologi of Cancer. Burgers Medicinal Chemistry and Drug Discovery Sixth
Edition, Volume 5: Chemotherapeutic Agents. Edited by Donald J. Abraham. ISBN 0-47137031-2 2003 John Wiley&Sons, Inc.

TUMOR SUPPRESSOR GENE

Retinoblastoma gene [RB1 gene]

rare form of childhood malignancy

forms: hereditary & sporadic

pRb

105-KDa nuclear protein

inhibits E2F [prevents G1 S transition]

inhibited by: phosphorylation viral

oncoproteins [E1A, HPV E7]

TUMOR SUPPRESSOR GENE

Tp53 gene

location: 17p13.1

product: p53 protein [53 KDa]

function: induces DNA repair or apoptosis

mutation: point mutation > deletion

results to: loss of function p53

Clinical conditions: carcinomas, Li Fraumeni Syndrome

p53 inhibited by: HPV E6 (genomic), microRNA(epigenetic)

Faulty proteins can interfere with normal

signal transduction pathways
Path producing a product that inhibits cell
division
Product of p53 tumor-suppressor gene is a
transcription factor
p53 transcription factor normally activates genes
for factors that stop cell division
In the absence of functional p53, cell division
continues because the inhibitory protein is not
produced (p21)

Cyclin Regulators
p 21: inhibits cell cycle progression and permits
DNA repair to take place.
P53: the guardian of the genome
In the presence of DNA damage, influences
transcription to either:
Halt cell cycle progression to facilitate DNA repair.
In cases of severe DNA damage, activates apoptosis.

Mutations in p53 are the most common genetic

alterations found in human cancer (50% of all human
tumors)

Growth Arrest

Video

Mechanisms of Oncogene Activation

1. Chrosomal
Mutation
2. DNA Mutation
3. Viral gene
integration

1. Chromosomal Mutation
a. Chromosomal Deletion
Ex: Retinoblastoma

1. Chromosomal Mutation
b. Chromosomal Amplification/ Duplication
Ex: Breast Ca, Cervical Ca, Lung Ca, Ovarian Ca, Hepatocellular
Ca, Esopagheal Ca, Colorectal Ca

1. Chromosomal Mutation
c. Chromosomal Inversion
Ex: Papillary thyroid carcinomas

1. Chromosomal Mutation
d. Chromosomal Translocation

d. Chromosomal Translocation

Ex. Burkitts Lymphoma

d. Chromosomal Translocation
Ex. Chronic Myelogenous Leukemia [CML]

2. DNA MUTATIONS

A change in the DNA sequence of the gene

Mutations can alter protein product of DNA, stop

gene working or activate gene

Sudden, random alteration of original DNA code

that changes the genotype.

Can be caused by chemical or environmental

factors - mutagens.

2. DNA MUTATIONS
Types of Mutation :
Deletion - DNA missing
Insertion - extra DNA inserted
Expansion - DNA repeat size has
increased
Point Mutation - change in one base

2. DNA MUTATIONS
Types of Mutation
AGC
AGC
AGC
AGC
AGC

(in coding sequence)

TTC GAC CCG

Wild type (Normal)
T_CG ACC CG
Deletion
TTC CGA CCC G Insertion
TTC TTC GAC CCG Expansion
TTC GAC CGG Point mutation

POINT MUTATION
UAA
(Termination Codon)
UCA
(Codon for Serine)
UCU
(Codon for Serine)
CCA
(Codon for Proline)

3. Viral Gene Integration

promoter

Viral promoter

Viral Carcinogenesis
Viral carcinogens are classified into
RNA and DNA viruses.
Most RNA oncogenic viruses belong
to the family of retroviruses that
contain reverse transcriptase
mediates transfer of viral RNA into
virus specific DNA.

Viruses Associated With The

Development Of Human
Neoplasia
VIRUSES

NEOPLASMS

DNA VIRUSES

Human papilloma virus

Cervical Ca, warts,
anogenital carcinoma
Herpes simplex virus II
Cervical
carcinoma
Epstein-Barr virus
NPCa, African
Burkitts
Human Herpes virus 8
Kaposis sarcoma
Hepatitis B virus
Hepatocellular Ca
Herpes simplex virus 6
Certain B cell
(HBLV)
lymphomas

Viruses Associated With The

Development Of Human
Neoplasia
VIRUSES
RNA VIRUSES

NEOPLASMS

Human T-cell leukemia virus I

leukemia,
lymphoma
Human T-cell leukemia virus II
of hairy

Some T-cell

Some cases
cell

leukemia
Human immunodeficiency virus I
Kaposis

Lymphoma;
sarcoma

VIRAL AGENTS: DNA

Human Papillomavirus [HPV viruses
types 16, 18, 31, 33 & 35]

E2 is not expressed (E2: repression of E6/E7 genes)

Over-expression
E6 (inhibits p53)
E7 (inhibits pRb)

Transformasi sel pejamu akibat insersi sebuah promotor

virus atau onkogen virus

CANCER CELLS
AND
NORMAL
CELLS

Molecular Biologi of Cancer. Burgers Medicinal Chemistry and Drug Discovery Sixth Edition, Volume
5: Chemotherapeutic Agents. Edited by Donald J. Abraham. ISBN 0-471-37031-2 2003 John
Wiley&Sons, Inc.

Molecular Biologi of Cancer. Burgers Medicinal Chemistry and Drug Discovery Sixth Edition, Volume
5: Chemotherapeutic Agents. Edited by Donald J. Abraham. ISBN 0-471-37031-2 2003 John
Wiley&Sons, Inc.

INVASION-INTRAVASATIONMETASTASIS
The defining characteristic of a malignancy.
Invasion: active translocation of neoplastic
cells across tissue barriers.
Critical pathologic point: local invasion and
neovascularization. These events may occur
before clinical detection.

ANGIOGENESIS
Formation of new blood vessels from
existing vascular bed
Carried out by endothelial cells (EC)
and extra cellular matrix (ECM)
Regulated by angiogenic factors
(inducers and inhibitors)
* A tumor is unable to grow larger
than 1 mm3 w/o developing a
new blood supply

ANGIOGENESIS
As tumor size increases, intratumoral
O2 levels fall and the center of the
mass becomes hypoxic leading to
up-regulation of the hypoxia inducible
factor (HIF1)
An important transcriptional target of
HIF1 is the VEGF growth factor,
induces neovascularization of tumors

INVASION-INTRAVASATIONMETASTASIS

D. and Zhao J. 2013. The Role of chemokine receptor CXCR4 in breast cancer metastasis. Am J Cancer Res 3(1); 46-57.

CARCINOGENS
Occupation related causes
Lifestyle related causes
Tobacco
Diet
Sexual practices

Viral carcinogens
Physical carcinogens
Chemical carcinogens

Occupational Risk Factors

Etiology
Arsenic
Asbestos
Benzene
Benzedine
Chromium cpds
Radiation (mining)
Mustard gas
Polycyclic hydrocarbons
Vinyl Chloride

Site of Malignancy
Lung, skin, liver
Mesothelium, lung
Leukemia
Bladder
Lung
Numerous locations
Lung
Lung, skin
Angiosarcoma of liver

Lifestyle Risk Factors

Tobacco-related:
Lung cancer
Pancreatic cancer
Bladder cancer
Renal cancer
Cervical cancer

Diet-Related Risk Factors

Salt
Low vitamins A, C,
E
Low consumption
of yellow-green
vegetables

Gastric
Cancer
Esophageal
Cancer

Diet-Related Risk Factors

High fat
Low fiber
Low calcium
High fried
foods

Mycotoxin
s

Colon Cancer
Pancreatic
Cancer
Prostate Cancer
Breast Cancer
Uterine Cancer

Liver Cancer

Sexual Practices Risk

Factors
Sexual promiscuity
Multiple partners
Unsafe Sex
Human
Papillomavirus

Cervical
Cancer

PHYSICAL CARCINOGENESIS
Radiation:

Ionizing - X-rays, rays, cosmic rays

Non-ionizing - UV light

Ionizing Radiation

includes electromagnetic rays & particulate matter

mechanism: free radicals & mutations

pathology: leukemia, thyroid ca, lung & breast ca

RADIATION

PRE-IRRADIATION

POST-IRRADIATION

PHYSICAL
CARCINOGENESIS
Ultraviolet Rays
UV-C filtered by ozone
UV-B
Inhibition of cell division
inactivation of enzymes
induction of mutations
cell death at high doses
Squamous cell cancer
Basal cell cancer
Melanocarcinoma

Efek radiasi atau karsinogen kimia pada

protoonkogen atau promotor. Mutasi dapat berupa
point of mutation, delesi atau insersi

DNA Mutation and Repair

Cells contain many DNA repair
mechanisms.
Example - exposure to UV light.
UV - High energy light.
- Causes formation of thymine
dimer.
- at low levels, it can be repaired.
- at high levels, it can be deadly.

Photodimerization

Exposure to UV light
can cause adjacent
thymines to
covalently link.
This results in a
distortion of the DNA
molecule and
breaks the hydrogen
bonding with the
Adenine

UV light
| | |
A C T

|
T

T
|

A C G T A
| | | | |

G A
| |

| | | |
G C A T

| |
A C

| | | |
G Cthymine
A T

T
|

C Gdimer
T A
| | | |

G
|

Nucleotide Excision
Repair (NER)
Separation

Incision

Operates by a cut-and patch

mechanism that removes a
variety of bulky lesions,
including pyrimidines dimers and
nucleotides which various
chemical groups have become
attached.

Excision

Ligation

Two pathways distinguished in

NER
- Transcription-coupled pathway
(repairs the template)
- Global pathway (corrects DNA
strands in the remainder of
the genome)

Nucleotide Excision Repair (NER)

Damage recognition in the global pathway is mediated

by an XPC protein whereas damage recognition in the
transcription-coupled pathways is thought to be
mediated by a stalled RNA polymerase in conjunction
with a CSB protein.

DNA strand separation (by XPB and XPD proteins, two

helicase subunits of TFIIH) (2)

Incision (by XPG on the 3side and the XPF-ERCC1

complex on the 5side) (3)

Excision (4)

DNA repair synthesis (by DNA polymerase and/or ) (5)

Ligation (by DNA ligase I).

CHEMICAL
CARCINOGENESIS
Carcinogens

cyclophosphamide

Promoters

saccharine & cyclamates

chlorambucil

Estrogen

busulfan

Diesthystilbestrol [DES]

melphalan

Base analogs

2-aminopurine
5-bromouracil

IMUNOLOGI TUMOR
Sel tumor berbeda dengan sel normal
dikenal sebagai non-self/foreign (antigen)
Respon imun gagal menghambat
pertumbuhan sel tumor
Antigen dikenal sebagai hasil :
Mutasi
Abnormal Expression
Oncogenic viruses
Oncofetal antigens
Altered surface modifications
Tissue specific differentiations

Olivera J. Finn, Ph.D. Cancer Immunology.N Engl J Med 2008;358:2704-15.

Tumor antigen, ada 2 tipe :

1. Tumor-Specific Antigens (TSA)/
Tumor-Specific Transplantation
Antigens (TSTAs)

Spesifik untuk tumor, tidak ada pada sel

normal
Hasil mutasi pada sel tumor yang
menghasilkan perubahan pada protein
seluler MHC kelas I induksi respon
imun dimediasi oleh CTL spesifik tumor
Protein yang dibentuk aibat mutasi 1
atau lebih gen

2. Tumor-Associated Antigens (TAAs)/

Tumor-Associated Transplantation
Antigens (TATAs)

Tidak spesifik untuk sel tumor

Respon Imun Terhadap Tumor

Ada 4 jenis sel-sel imun yang berbeda dapat
membunuh target tumor in vitro maupun in vivo :
1. Sel pembunuh alami (Natural Killer / NK Cell)
2. Cytolytic thymus dependent Lymphocytes
(CTLs)
3. Lymphokine-activated killer cells (LAK cells)
4. Macrophages

1. Natural Killer (NK) Cell

Sel-sel NK dapat membunuh sel-sel tumor tanpa
mensintesa sebelumnya antigen spesifik,
aktivitasnya tidak memerlukan adanya MHC kelas I
pada sel-sel target.
NK Berkembang dalam bone marrow, kemudian
diperoleh dalam peripheral blood, sel pit (sinusoid
liver) dan sinusoid limpa
Dapat mensekresi interferon gamma, dan secara
spontan membunuh sel yang diinfeksi virus dan selsel tumor
Sel NK ambil bagian dalam pengawasan tumor yang
mulai timbul dan juga terhadap pertumbuhan
metastatik tumor.

Aktivasi Sel Natural Killer

2. CYTOLYTIC THYMUS-DEPENDENT
LYMPHOCYTES (CTLs) = Cytotoxic T cells

CTLs dapat membunuh tumor setelah

dipresentasikan oleh MHC kelas I
CTLs dapat membunuh sel-sel target
melalui jalur yang memerlukan crosslinked ligands CTLs dengan reseptor
pada permukaan spesifik sel-sel tumor
untuk merangsang apoptosis sel-sel
tumor (program kematian sel).

AKTIVASI CYTOLYTIC THYMUS-DEPENDENT

LYMPHOCYTES (CTLs) = Cytotoxic T cells

Daniel S. Chen and Ira Mellman, Oncology Meets Immunology: The Cancer-Immunity
Cycle. Immunity 39, July 25, 2013. Elsevier Inc.

3. Lymphokine-Activated Killer
cells (LAK cells)
Subset null lymphocyte berbeda
dari sel-sel NK dan CTLs
Dapat dihasilkan in vitro, caranya
mengkultur sel-sel limfosit +
Interleukin 2, konsentrasi IL2 tinggi
Memiliki aktivitas anti tumor
(membunuh sel tumor).

Sistem Imun Tubuh

4. Macrophages
Sel fagosit mononukleus non limfosit
Ada pada jaringan dan dalam darah,
derivat dari stem sel monositic.
Penting sebagai sel pelengkap pada
respon imun
Makrofag khusus ada pada beberapa
lokasi, sel-sel Kupffer dan histiosit.

4. Macrophages
Diaktifkan sebagai suatu hasil dari reaksi delayed
Hypersentivity oleh sel T atau oleh aktivator
makrofag non spesifik, polinukleotida.
Peran: Sebagai scavengers , clean up injured
cells
Dapat diaktifkan oleh berbagai agents,
termasuk peptida bakteri seperti: mycobacteria
(BCG) dan polinukleotida yang dianggap anntigen
oleh tubuh
Mekanisme: membunuh dengan cara memfagosit,
melalui pembentukan oksigen radikal dan aktivasi
enzim proteolitik

Daniel S. Chen and Ira Mellman, Oncology Meets Immunology: The Cancer-Immunity
Cycle. Immunity 39, July 25, 2013. Elsevier Inc.

Daniel S. Chen and Ira Mellman, Oncology Meets Immunology: The Cancer-Immunity
Cycle. Immunity 39, July 25, 2013. Elsevier Inc.